117 research outputs found
Kinase Inhibitor Profile For Human Nek1, Nek6, And Nek7 And Analysis Of The Structural Basis For Inhibitor Specificity
Human Neks are a conserved protein kinase family related to cell cycle progression and cell division and are considered potential drug targets for the treatment of cancer and other pathologies. We screened the activation loop mutant kinases hNek1 and hNek2, wild-type hNek7, and five hNek6 variants in different activation/phosphorylation statesand compared them against 85 compounds using thermal shift denaturation. We identified three compounds with significant Tm shifts: JNK Inhibitor II for hNek1(Ä262-1258)-(T162A), Isogranulatimide for hNek6(S206A), and GSK-3 Inhibitor XIII for hNek7wt. Each one of these compounds was also validated by reducing the kinases activity by at least 25%. The binding sites for these compounds were identified by in silico docking at the ATP-binding site of the respective hNeks. Potential inhibitors were first screened by thermal shift assays, had their efficiency tested by a kinase assay, and were finally analyzed by molecular docking. Our findings corroborate the idea of ATP-competitive inhibition for hNek1 and hNek6 and suggest a novel non-competitive inhibition for hNek7 in regard to GSK-3 Inhibitor XIII. Our results demonstrate that our approach is useful for finding promising general and specific hNekscandidate inhibitors, which may also function as scaffolds to design more potent and selective inhibitors.20111761191Rubin, G.M., Yandell, M.D., Wortman, J.R., Gabor Miklos, G.L., Nelson, C.R., Hariharan, I.K., Fortini, M.E., Fleischmann, W., Comparative genomics of the eukaryotes (2000) Science, 287, pp. 2204-2215Johnson, L.N., Lowe, E.D., Noble, M.E., Owen, D.J., The Eleventh Datta Lecture. The structural basis for substrate recognition and control by protein kinases (1998) FEBS Lett., 430, pp. 1-11Hanks, S.K., Eukaryotic protein kinases (1991) Curr. Opin. Struct. Biol., 1, pp. 369-383Jeffrey, P.D., Russo, A.A., Polyak, K., Gibbs, E., Hurwitz, J., Massagué, J., Pavletich, N.P., Mechanism of CDK activation revealed by the structure of a cyclinA-CDK2 complex (1995) Nature, 376, pp. 313-320Yamaguchi, H., Hendrickson, W.A., Structural basis for activation of human lymphocyte kinase Lck upon tyrosine phosphorylation (1996) Nature, 384, pp. 484-489Canagarajah, B.J., Khokhlatchev, A., Cobb, M.H., Goldsmith, E.J., Activation mechanism of the MAP kinase ERK2 by dual phosphorylation (1997) Cell, 90, pp. 859-869Hubbard, S.R., Crystal structure of the activated insulin receptor tyrosine kinase in complex with peptide substrate and ATP analog (1997) EMBO J, 16, pp. 5572-5581Fry, A.M., O'Regan, L., Sabir, S.R., Bayliss, R., Cell cycle regulation by the NEK family of protein kinases (2012) J. Cell Sci., 125, pp. 4423-4433Meirelles, G.V., Perez, A.M., Souza, E.E., Basei, F.L., Papa, P.F., Melo Hanchuk, T.D., Cardoso, V.B., Kobarg, J., "Stop Ne(c)king around": How systems biology can help to characterize the functions of Nek family kinases from cell cycle regulation to DNA damage response (2014) World J. Biol. Chem., 5, pp. 141-160Fry, A.M., Mayor, T., Meraldi, P., Stierhof, Y.D., Tanaka, K., Nigg, E.A., C-Nap1, a novel centrosomal coiled-coil protein and candidate substrate of the cell cycle-regulated protein kinase Nek2 (1998) J. Cell Biol., 141, pp. 1563-1574Quarmby, L.M., Mahjoub, M.R., Caught nek-ing: Cilia and centrioles (2005) J. Cell Sci., 118, pp. 5161-5169Meirelles, G.V., Silva, J.C., Mendonça, Y.A., Ramos, C.H., Torriani, I.L., Kobarg, J., Human Nek6 is a monomeric mostly globular kinase with an unfolded short N-terminal domain (2011) BMC Struct. Biol., 11, p. 12Belham, C., Roig, J., Caldwell, J.A., Aoyama, Y., Kemp, B.E., Comb, M., Avruch, J., A mitotic cascade of NIMA family kinases. Nercc1/Nek9 activates the Nek6 and Nek7 kinases (2003) J. Biol. Chem., 278, pp. 34897-34909Yin, M.J., Shao, L., Voehringer, D., Smeal, T., Jallal, B., The serine/threonine kinase Nek6 is required for cell cycle progression through mitosis (2003) J. Biol. Chem., 278, pp. 52454-52460Yissachar, N., Salem, H., Tennenbaum, T., Motro, B., Nek7 kinase is enriched at the centrosome, and is required for proper spindle assembly and mitotic progression (2006) FEBS Lett., 580, pp. 6489-6495Kim, S., Lee, K., Rhee, K., NEK7 is a centrosomal kinase critical for microtubule nucleation (2007) Biochem. Biophys. Res.Commun., 360, pp. 56-62Roig, J., Mikhailov, A., Belham, C., Avruch, J., Nercc1, a mammalian NIMA-family kinase, binds the Ran GTPase and regulates mitotic progression (2002) Genes Dev., 16, pp. 1640-1658Upadhya, P., Birkenmeier, E.H., Birkenmeier, C.S., Barker, J.E., Mutations in a NIMA-related kinase gene, Nek1, cause pleiotropic effects including a progressive polycystic kidney disease in mice (2000) Proc. Natl. Acad. Sci. USA, 97, pp. 217-221Liu, S., Lu, W., Obara, T., Kuida, S., Lehoczky, J., Dewar, K., Drummond, I.A., Beier, D.R., A defect in a novel Nek-family kinase causes cystic kidney disease in the mouse and in zebrafish (2002) Development, 129, pp. 5839-5846Chen, J., Li, L., Zhang, Y., Yang, H., Wei, Y., Zhang, L., Liu, X., Yu, L., Interaction of Pin1 with Nek6 and characterization of their expression correlation in Chinese hepatocellular carcinoma patients (2006) Biochem. Biophys. Res. Commun., 341, pp. 1059-1065Chen, Y., Chen, P.L., Chen, C.F., Jiang, X., Riley, D.J., Never-in mitosis related kinase 1 functions in DNA damage response and checkpoint control (2008) Cell Cycle, 7, pp. 3194-3201Lee, M.Y., Kim, H.J., Kim, M.A., Jee, H.J., Kim, A.J., Bae, Y.S., Park, J.I., Yun, J., Nek6 is involved in G2/M phase cell cycle arrest through DNA damage induced phosphorylation (2008) Cell Cycle, 7, pp. 2705-2709Innocenti, P., Cheung, K.M., Solanki, S., Mas-Droux, C., Rowan, F., Yeoh, S., Boxall, K., Hardy, T., Design of potent and selective hybrid inhibitors of the mitotic kinase Nek2: Structure-activity relationship, structural biology, and cellular activity (2012) J. Med. Chem., 55, pp. 3228-3241Solanki, S., Innocenti, P., Mas-Droux, C., Boxall, K., Barillari, C., Van Montfort, R.L., Aherne, G.W., Hoelder, S., Benzimidazole inhibitors induce a DFG-out conformation of never in mitosis gene A-related kinase 2 (Nek2) without binding to the back pocket and reveal a nonlinear structure-activity relationship (2011) J. Med. Chem., 54, pp. 1626-1639Whelligan, D.K., Solanki, S., Taylor, D., Thomson, D.W., Cheung, K.M., Boxall, K., Mas-Droux, C., Grummitt, C.G., Aminopyrazine inhibitors binding to an unusual inactive conformation of the mitotic kinase Nek2: SAR and structural characterization (2010) J. Med. Chem., 53, pp. 7682-7698Srinivasan, P., ChellaPerumal, P., Sudha, A., Discovery of novel inhibitors for Nek6 protein through homology model assisted structure based virtual screening and molecular docking approaches (2014) Sci. World J., 2014. , ID: 967873Rellos, P., Ivins, F.J., Baxter, J.E., Pike, A., Nott, T.J., Parkinson, D.M., Das, S., Shen, Q.Y., Structure and regulation of the human Nek2 centrosomal kinase (2007) J. Biol. Chem., 282, pp. 6833-6842Westwood, I., Cheary, D.M., Baxter, J.E., Richards, M.W., Van Montfort, R.L., Fry, A.M., Bayliss, R., Insights into the conformational variability and regulation of human Nek2 kinase (2009) J. Mol. Biol, 386, pp. 476-485Richards, M.W., O'Regan, L., Mas-Droux, C., Blot, J.M., Cheung, J., Hoelder, S., Fry, A.M., Bayliss, R., An autoinhibitory tyrosine motif in the cell-cycleregulated Nek7 kinase is released through binding of Nek9 (2009) Mol. Cell, 36, pp. 560-570Geromichalos, G.D., Importance of molecular computer modeling in anticancer drug development (2007) J. Buon., 12, pp. S101-S118Vedadi, M., Niesen, F.H., Allali-Hassani, A., Fedorov, O.Y., Finerty, P.J., Jr., Wasney, G.A., Yeung, R., Berglund, H., Chemical screening methods to identify ligands that promote protein stability, protein crystallization, and structure determination (2006) Proc. Natl. Acad. Sci. USA, 103, pp. 15835-15840Lengauer, T., Rarey, M., Computational methods for biomolecular docking (1996) Curr. Opin. Struct. Biol., 6, pp. 402-406Fedorov, O., Marsden, B., Pogacic, V., Rellos, P., Müller, S., Bullock, A.N., Schwaller, J., Knapp, S., A systematic interaction map of validated kinase inhibitors with Ser/Thr kinases (2007) Proc. Natl. Acad. Sci. USA, 104, pp. 20523-20528Roberge, M., Berlinck, R.G., Xu, L., Anderson, H.J., Lim, L.Y., Curman, D., Stringer, C.M., Vincent, I., High-throughput assay for G2 checkpoint inhibitors and identification of the structurally novel compound isogranulatimide (1998) Cancer Res, 58, pp. 5701-5706Jiang, X., Zhao, B., Britton, R., Lim, L.Y., Leong, D., Sanghera, J.S., Zhou, B.B., Roberge, M., Inhibition of Chk1 by the G2 DNA damage checkpoint inhibitor isogranulatimide (2004) Mol. Cancer Ther., 3, pp. 1221-1227De Castro, E., Sigrist, C.J.A., Gattiker, A., Bulliard, V., Langendijk-Genevaux, P.S., Gasteiger, E., Bairoch, A., Hulo, N., ScanProsite: Detection of PROSITE signature matches and ProRule-associated functional and structural residues in proteins (2006) Nucleic Acids Res., 34, pp. W362-W365Gavrin, L.K., Saiah, E., Approaches to discover non-ATP site kinase inhibitors (2013) Med. Chem. Commun., 4, pp. 41-51Oliveira, S.H., Ferraz, F.A., Honorato, R.V., Xavier-Neto, J., Sobreira, T.J., De Oliveira, P.S., KVFinder: Steered identification of protein cavities as a PyMOL plugin (2014) BMC Bioinform., 15, p. 197Meirelles, G.V., Lanza, D.C.F., Silva, J.C., Bernachi, J.S., Leme, A.F.P., Kobarg, J., Characterization of hNek6 Interactome Reveals an Important Role for Its Short N-Terminal Domain and Colocalization with Proteins at the Centrosome (2010) J. Proteome Res., 9, pp. 6298-6316Trott, O., Olson, A.J., AutoDockVina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization and multithreading (2010) J. Comput. Chem., 31, pp. 455-46
Comportamento de dois genótipos de milho cultivados em sistema de aléias preestabelecido com diferentes leguminosas arbóreas.
O cultivo em aléias tem sido recomendado como alternativa para a substituição da agricultura de corte e queima, no trópico úmido, devido à grande capacidade de produção de matéria orgânica e de reciclagem de nutrientes, mas algumas dúvidas quanto à sustentabilidade e à competição interespecífica são persistentes. O objetivo no trabalho foi avaliar a viabilidade da cultura do milho em um sistema de cultivo em aléias de leguminosas arbóreas. O delineamento experimental utilizado foi em blocos casualisados, com quatro repetições dos tratamentos: aléias de sombreiro (Clitoria fairchildiana), ingá (Inga edulis), guandu (Cajanus cajan) e leucena (Leucaena leucocephala) e uma testemunha sem aléias. Foram avaliadas a remobilização de carbono e nitrogênio, massa de grãos, massa de mil grãos e competição interespecífica entre as cultivares de milho e as leguminosas. A produção de grãos foi maior nas parcelas com C. fairchildiana e L. leucocephala. A produtividade do híbrido de milho foi superior à da variedade em todos os tratamentos. A produtividade e a massa de mil grãos de milho não são negativamente afetadas pela distância da linha da leguminosa arbórea. Esse estudo conclui que o sistema de aléias com leguminosas arbóreas é uma alternativa importante ao manejo sustentável dos agroecossistemas no tropico úmido. Além disso, nessa região a produtividade em grãos na cultura do milho é favorecida no sistema de aléias preeestabelecidas com as leguminosas arbóreas sombreiro, ingá e leucena e pela utilização de genótipos eficientes no aproveitamento do nitrogênio, cujo sincronismo entre a liberação e a absorção do N aplicado por meio das leguminosas deve ser aprimorado
Avaliação do módulo de elasticidade em concreto com metacaulim de alta reatividade, proveniente de rejeito industrial
Updated precision measurement of the average lifetime of B hadrons
The measurement of the average lifetime of B hadrons using inclusively reconstructed secondary vertices has been updated using both an improved processing of previous data and additional statistics from new data. This has reduced the statistical and systematic uncertainties and gives \tau_{\mathrm{B}} = 1.582 \pm 0.011\ \mathrm{(stat.)} \pm 0.027\ \mathrm{(syst.)}\ \mathrm{ps.} Combining this result with the previous result based on charged particle impact parameter distributions yields \tau_{\mathrm{B}} = 1.575 \pm 0.010\ \mathrm{(stat.)} \pm 0.026\ \mathrm{(syst.)}\ \mathrm{ps.
Limits on the production of scalar leptoquarks from Z (0) decays at LEP
A search has been made for pairs and for single production of scalar leptoquarks of the first and second generations using a data sample of 392000 Z0 decays from the DELPHI detector at LEP 1. No signal was found and limits on the leptoquark mass, production cross section and branching ratio were set. A mass limit at 95% confidence level of 45.5 GeV/c2 was obtained for leptoquark pair production. The search for the production of a single leptoquark probed the mass region above this limit and its results exclude first and second generation leptoquarks D0 with masses below 65 GeV/c2 and 73 GeV/c2 respectively, at 95% confidence level, assuming that the D0lq Yukawa coupling alpha(lambda) is equal to the electromagnetic one. An upper limit is also given on the coupling alpha(lambda) as a function of the leptoquark mass m(D0)
Caracterização físico-química de polpas de frutos da Amazônia e sua correlação com a atividade anti-radical livre
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